tetraesters from the esterification of pentaerythritol with pentanoic acid, heptanoic acid and 3,5,5-trimethylhexanoic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

May 06th, 1996 - August 29th, 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study, tested with the source substanceCAS 11138-60-6. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate

EC Number:

234-392-1

EC Name:

Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate

Cas Number:

11138-60-6

IUPAC Name:

2,2-bis[(octanoyloxy)methyl]butyl decanoate

Details on test material:

- Physical state: Yellow , clear liquid
- Expiration date: 01.01.2006
- Storage condition of test material: room temperature, protected from light

Method

Target gene:

Genes involved in Histidine synthesis

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix (Spraque Dawley rats, male, Aroclor 1254 induced)

Test concentrations with justification for top dose:

0, 10, 33, 100, 333 and 1000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: see table

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h
- Expression time (cells in growth medium): 48 to 72 h

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial backgroung lawn wit a dissecting microscope

Evaluation criteria:

Revertant colonies were counted and the mean and standard deviation were calculated and compared to the controls.
All Salmonella tester strains must demonstrate the presence of the deep rough mutation and the deletion of the uvrA gene. Cultures of the TA98 and TA100 strains must demonstrate the presence of the pKM101 plasmid R-factor. All WP2 uvrA cultures must demonstrate the deletion of the uvrA gene. All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls. Tester strain titers must be above 30.000.000 cells/ml. The mean of each positive control must be at least three-fold increased to the controlls. A minimum of three non-toxic dose levels are recquired to evaluate assay data.

Statistics:

Mean and standard deviation were calculated

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

other: An unacceptable vehicle control value with the tester strain TA1537, contamination of tester strain TA98, experiments were repeated

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: >100 µg/plate

RANGE-FINDING/SCREENING STUDIES: Yes

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Experiment 1:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 1535	TA1537	TA98	TA100	TA 1538	WP2uvrA
-	Vehicle	12	4	17	111	5	21
	10	3	4	15	115	6	16
	33	5	5	19	113	6	11
-	100	9	4	19	98	7	12
-	333	7	5	11	116	4	10
-	1000	8	6	21	120	7	13
Positive controls - S9	Name	SA	9AA	2NF	SA	2NF	MMS
	Concentrations (μg/plate)	1.0	75	1.0	1.0	1.0	1000
	Number of colonies/plate	241	40	105	377	179	143
+	Vehicle	14	5	18	133	7	11
	10	9	5	27	114	12	16
+	33	9	3	21	113	8	13
+	100	8	7	26	108	9	13
+	333	9	4	27	115	6	8
+	1000	10	5	17	111	9	13
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1.0	1.0	1.0	1.0	1.0	10
	Number of colonies/plate	72	127	888	904	783	57

SA: Sodium azide

9AA : 9-Aminoacridine

MMS: Methylmethanesulfonate

2-AA: 2-Aminoanthracene

2NF: 2-Nitrofluorene

 

 

Experiment 2/3:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 1535	TA1537	TA98	TA100	TA 1538	WP2uvrA
-	Vehicle	14	10	23	126	11	27
	10	7	13	16	117	7	27
	33	9	15	23	124	8	19
-	100	5	13	22	120	6	18
-	333	12	9	17	110	11	16
-	1000	8	14	17	125	5	21
Positive controls - S9	Name	SA	9AA	2NF	SA	2NF	MMS
	Concentrations (μg/plate)	1.0	75	1.0	1.0	1.0	1000
	Number of colonies/plate	429	757	125	601	221	195
+	Vehicle	8	5	19	147	14	24
	10	9	7	17	142	16	30
+	33	10	5	19	136	18	25
+	100	10	5	20	132	13	31
+	333	10	4	17	138	12	19
+	1000	10	7	19	125	12	19
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1.0	1.0	1.0	1.0	1.0	10
	Number of colonies/plate	85	97	530	647	1041	88

SA: Sodium azide

9AA : 9-Aminoacridine

MMS: Methylmethanesulfonate

2-AA: 2-Aminoanthracene

2NF: 2-Nitrofluorene

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative Under the tested experimental conditions the test substance did not induce gene mutations in S. typhimurium strains and in an E. coli strain up to the maximum of solubility. Therefore it is not considered to be mutagenic in this bacterial mutagenicity tes